You don’t mess with Jackie Hoffman. At least, not if you’re going to ask her to perform at your new nightspot.

Kicking off her show at 54 Below, the comedienne milked her chagrin at being the first entertainer to play the new club, only to see Patti LuPone officially christen the joint days later.

In her typical take-no-prisoners style, Hoffman took generous potshots at LuPone, “Smash” (whose Scott Wittman is the club’s creative consultant) and the subterranean venue itself.

“Gays love the bottom, and that’s where I am,” she sings in the opening number, “Bottom,” before adding, “Boy, if this ceiling could talk.”

But the self-styled diva’s biggest target is herself. For this gig, co-written and directed by Michael Schiralli, she laments her standing as No. 33 in a magazine’s list of the 50 funniest New Yorkers. She also delivers a medley of her minimal musical contributions to the Broadway shows “Hairspray,” “Xanadu” and “The Addams Family” — the latter consisting entirely of finger snaps.

She kvetches over such missed career opportunities as a failed audition for a Woody Allen play — funny, she seems like perfect casting to me —and not even being allowed to audition for Miss Hannigan in the upcoming “Annie” revival.

Singing mostly original songs, many with music by her very young-looking pianist, Will Van Dyke, she’s one of the few performers who can get away with singing a humorous ditty about autism. While strumming a ukulele, no less.

Her self-deprecation knows no bounds. Looking smashing in a form-fitting black dress, she insists her career has nowhere to go but down.